sie-exclusion chromatography columns and tandards · 2018-05-28 · size-exclusion chromatography...

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Size-Exclusion Chromatography Colum

ns and Standards

Size-Exclusion Chromatography Columns and Standards

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Contents

GPC Columns for Non-Aqueous Samples ......................................................................................301

ACQUITY APC XT Columns...................................................................................................................................301

Styragel Columns Selection Guide ............................................................................................................... 303

Ultrastyragel Columns .............................................................................................................................................307

HSPgel Columns ...........................................................................................................................................................307

Shodex Columns ............................................................................................................................................................310

Envirogel High-Resolution GPC Cleanup Columns............................................................................311

SEC Columns ...................................................................................................................................................312

ACQUITY APC AQ Columns .................................................................................................................................312

Ultrahydrogel Columns ............................................................................................................................................314

HSPgel Columns ............................................................................................................................................................315

ACQUITY UPLC Protein SEC Columns ........................................................................................................315

XBridge Protein BEH SEC Columns ...............................................................................................................316

Protein Standards ......................................................................................................................................................... 317

Solvent Guide ..................................................................................................................................................318

Calibration Standards ...............................................................................................................................320

ACQUITY APC Calibration Standards ..........................................................................................................320

ReadyCal Standards ...................................................................................................................................................321

Polymer Specific Calibration Standards ...................................................................................................322

Individual MW Standards ......................................................................................................................................323

SEC Calibration Standards ...................................................................................................................................323

301

Size-Exclusion Chromatography Columns and StandardsFor over 50 years, Waters has continuously improved GPC (Gel Permeation Chromatography), and SEC (Size Exclusion Chromatography), refining instrumentation, packing materials, and technology. Among the resultant innovations are size-exclusion techniques that expand beyond the original polymer analysis. These include applications for separating small and large molecules from interfering matrices such as those in foods, pharmaceutical preparations, and natural products.

As a market leader and a primary manufacturer of chromatographic instrumentation and consumables, Waters will continue to influence the field of separation science, providing the highest quality products and expert applications support.

GPC Columns for Non-Aqueous SamplesThe goals for a separation can range between maximum speed, for screening purposes, to maximum resolution, for quality control purposes. Each analysis type presents unique challenges. Waters’ comprehensive line of GPC columns ensures that the column or column bank you select for an analysis will accommodate a particular temperature, solvent, and polymer type.

ACQUITY APC XT COLUMNSUsing ACQUITY APC XT Columns, you can quantify and characterize polymer samples with accuracy and confidence while maximizing productivity. The high-performance chemistries contained in ACQUITY Advanced Polymer Chromatography® (APC™) Columns enable rapid and accurate chromatographic characterization of synthetic polymer and macromolecular species. The rigid hybrid particles used for ACQUITY APC XT Columns provide an unprecedented capability for rapid solvent switching, allowing the scientist to use multiple conditions for the same column bank. This gives the chromatographer the ability to quantify and characterize polymer samples with confidence and accuracy while maximizing productivity.

Compared with conventional columns, ACQUITY APC Columns provide faster analysis time and increase chromatographic resolution. Improving data quality enhances your ability to accurately characterize polymers and to do it with confidence. The conventional GPC separation was performed using three Styragel® HR Columns (HR 0.5, HR 2, and HR 4E), all 7.8 × 300 mm. The same polystyrene sample was analyzed using a 3-column bank of 4.6 × 150 mm ACQUITY APC Columns (XT 45, XT 45, and XT 200). The separation used THF; the flow rate was 1 mL/min.

MV

0

5

10

15

20

25

30

35

13.0 14.0 15.0 16.0 17.0 18.0 19.0 20.0 21.0 22.0 23.0 24.0 25.0 26 min

100K

10K

1K

Conventional GPC System3 × Styragel 7.8 × 300 mm

µRIU

0

2

4

6

8

10

12

14

16

18

20

2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.7 min

100K

10K

1K

ACQUITY APC System3 × ACQUITY APC Columns 4.6 × 150 mm

302 www.waters.com/orderwww.waters.com/gpccolumns

10 101 102 103 104 105 106 107

Effective Molecular Weight Range

ACQUITY APC XT 45

ACQUITY APC XT 125

ACQUITY APC XT 200

ACQUITY APC XT 450

ACQUITY APC XT 900Column:

ACQUITY APC XT Column Selection Guide

100

1,000

10,000

100,000

1,000,000

10,000,000

0.50 1.00 1.50 2.00

Pol

ysty

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Mol

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Elution volume (mL)

ACQUITY APC XT 450

ACQUITY APC XT 200

ACQUITY APC XT 900

ACQUITY APC XT 125

ACQUITY APC XT 45

Polystyrene calibration curves for ACQUITY APC XT Columns.

Ordering Information

ACQUITY APC XT Columns

P/N P/N P/N

Pore Size Effective MW Range* Particle SizeColumn Length

30 mm 75 mm 150 mm

45Å 200–5000 1.7 µm 186006992 186006993 186006995

125Å 1000–30,000 2.5 µm 186006997 186006998 186007000

200Å 3000–70,000 2.5 µm 186007002 186007003 186007005

450Å 20,000–400,000 2.5 µm 186007007 186007008 186007010

900Å 300,000–2,000,000 2.5 µm 186007252 186007253 186007254

* The calibration range is based on well-characterized polystyrene standards. All columns are 4.6 mm I.D., maximum temperature limit 90 ˚C and are shipped dry.

ACQUITY APC XT Columns are shipped dry, with acetal compression plugs at the assembly’s ends. If you are storing the columns wet using a strong solvating solvent, consider fitting compression plugs made of stainless steel.

ACQUITY APC XT Fitting Compression Plug

Description P/N

Stainless Steel Pin Plug, 1/16 in., High Pressure, 5/pk 700002747

303

Waters ACQUITY APC Column Selector

Easily find column and calibration kit recommendations that fit your polymer analysis requirements.

To try this tool, go to www.waters.com/APCselector

STYRAGEL COLUMNS SELECTION GUIDEWaters offers a comprehensive selection of polymeric GPC Columns. Select a column or column bank that is compatible with the temperature, solvent, and polymer type analyzed. Refer to the following charts to quickly compare the molecular weight ranges for the specified columns. By connecting two or more columns in series, you extend the effective molecular-weight range, which is necessary preparation for performing increasingly complex sample analyses.

101 102 103 104 105 106 107 108Effective Molecular Weight Range


UHDPEUH-PolystyreneUH-IsopreneUH-PMMA

Ultra-High Molecular Weight Waters Styragel HMW Series20 µm particle sizeAmbient to 150 oC

Styragel HMW 7

Styragel HMW 6E

Styragel HMW 2

Applications:

ABSAcetylAcrylicsEVANylonPEEKPET/PBT

Mid-Range Molecular WeightWaters Styragel HT Series10 µm particle sizeAmbient to 150 oC

Styragel HT 6E

Styragel HT 6

Styragel HT 5

Styragel HT 4

Styragel HT 3

Styragel HT 2

Applications:

AdditivesEpoxyOligomersPhenolicsUnsaturated PolyesterUrea/FormaldehydePolyethylene GlycolEthanolaminesMelamine Resin

Styragel HR 5E

Styragel HR 4E

Styragel HR 6

Styragel HR 5

Styragel HR 4

Styragel HR 3

Styragel HR 2

Styragel HR 1

Styragel HR 0.5

Low Molecular Weight Waters Styragel HR Series5 µm particle sizeAmbient to 80 oC

Applications:

101 102 103 104 105 106 107 108

Selection Guide

HMW—High Molecular Weight HT—High Temperature HR—High Resolution


Styragel HR (High-Resolution) Columns

Designed especially for low-molecular-weight samples, Waters Styragel HR Columns are ideal for analyzing oligomers, epoxies, and polymer additives, where high resolution is critical. Packed with rigid 5 µm particles, these columns deliver unrivaled resolution and efficiency in the low-to-mid molecular-weight region.

Calibration Curves for the Waters Styragel HR Series of High-Resolution Columns

Styragel HR

Mol

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12 min10864102

103

104

105

106

107

Styragel HR 0.5Styragel HR 1Styragel HR 2Styragel HR 3

Styragel HR 4Styragel HR 5Styragel HR 6Styragel HR 4EStyragel HR 5E

Conditions: Sample: PolystyreneMobile phase: THFFlow rate: 1 mL/min

45 min403530252045 min4035302520

45 min403530252045 min4035302520

Conventional Styragel HR Columns 7.8 × 300 mm:Column bank: Styragel HR 0.5, 1, 2, and 3Part numbers: WAT044231, WAT044234, WAT044237, WAT044222Mobile phase: THFColumn temp.: 35 °CFlow rate: 1 mL/minSample: Polystyrene standards: 0.5 K, 5.05 K, 49.8 K

Solvent-Efficient Styragel HR Columns 4.6 × 300 mm:Column bank: Styragel HR 0.5, 1, 2, and 3Part numbers: WAT045835, WAT045850, WAT045865, WAT045880Mobile phase: THFColumn temp.: 35 °CFlow rate: 0.35 mL/minSample: Polystyrene standard mix 0.5 K, 5.05 K, 49.8 K

Styragel HR Columns for Unrivaled Resolution of Low Molecular Weight Samples


Styragel HR Columns (7.8 × 300 mm)

P/N P/N P/N

Description Effective MW Range THF DMF Toluene

Styragel HR 0.5 0–1000 WAT044231 WAT044232 WAT044230

Styragel HR 1 100–5000 WAT044234 WAT044235 WAT044233

Styragel HR 2 500–20,000 WAT044237 WAT044238 WAT044236



Styragel HR 4E 50–100,000 WAT044240 WAT044241 WAT044239

Styragel HR 5 50,000–4,000,000 WAT054460 WAT054466 WAT054464

Styragel HR 5E 2000–4,000,000 WAT044228 WAT044229 WAT044227

Styragel HR 6 200,000–10,000,000 WAT054468 WAT054474 WAT054470

Styragel Guard Column 4.6 × 30 mm — WAT054405 WAT054415 WAT054410

305

Styragel HR Columns (4.6 × 300 mm)*

P/N P/N P/N


Styragel HR 0.5 0–1000 WAT045835 WAT045840 WAT045830

Styragel HR 1 100–5000 WAT045850 WAT045855 WAT045845




Styragel HR 4E 50–100,000 WAT045805 WAT045810 WAT045800

Styragel HR 5E 2000–4,000,000 WAT045820 WAT045825 WAT045815

*The same high performance as our conventional Styragel HMW Columns with the added advantage of reducing your solvent consumption by two-thirds.

Styragel HT (High-Temperature) Columns

You can use Styragel HT Columns with aggressive solvents at high temperatures without sacrificing resolution or column lifetime. Packed with rigid 10 µm particles, a typical plate count exceeds 10,000 plates per column. These columns are extremely durable because of a narrow particle-size distribution that results in a stable column bed. Suitable for both ambient and high-temperature analysis, the Styragel HT Columns offer excellent resolution of polymers in the mid-to-high molecular-weight range.

Styragel HT Columns Deliver Superior Performance— Even at High Temperatures

Branching Cumulative Distribution

Distribution

3 4 5 6Log (MW)

% M

W

100

80

60

40

20

0

Conditions: Column: Styragel HT 6E, 10 µm, 7.8 × 300 mmPart number: WAT044218Mobile phase: TCBColumn temp.: 140 °CSample: LDPE

Calibration Curves for the Waters Styragel HT Series of High-Temperature Columns

Styragel HMW

Mol

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12 min10 117 8 94 5 6102

103

104

105

106

107

108

109

Conditions: Sample: PolystyreneMobile phase: THFFlow rate: 1 mL/min


Styragel HT Columns (7.8 × 300 mm)

P/N P/N P/N


Styragel HT 2 100–10,000 WAT054475 WAT054480 WAT054476



Styragel HT 5 50,000–4,000,000 WAT044213 WAT044214 WAT044212


Styragel HT 6E 5000–10,000,000 WAT044219 WAT044220 WAT044218



Styragel HT Columns (4.6 × 300 mm)*

P/N P/N P/N






Styragel HT 6E 5000–10,000,000 WAT045980 WAT045985 WAT045975

*The same high performance as our conventional Styragel HT Columns with the added advantage of reducing your solvent consumption by two-thirds.

Styragel HMW (High-Molecular Weight) Columns

The Styragel HMW Columns are designed specifically to analyze polymers of ultra-high molecular-weight, which are susceptible to shearing. Combining high-porosity, 10 µm frits and 20 µm particles, the Styragel HMW Columns minimize polymer shear effects. Usable at ambient or elevated temperatures, these state-of-the-art columns exhibit excellent lifetimes.


Styragel HMW Columns (7.8 × 300 mm)

P/N P/N P/N


Styragel HMW 2 100–10,000 WAT054488 WAT054494 WAT054490

Styragel HMW 7 500,000–1 × 108 WAT044201 WAT044202 WAT044200

Styragel HMW 6E 5000–1 × 107 WAT044204 WAT044205 WAT044203


Styragel HMW Columns (4.6 × 300 mm)*

P/N P/N P/N


Styragel HMW 7 500,000–1 × 108 WAT046805 WAT046810 WAT046800

Styragel HMW 6E 5000–1 × 107 WAT046820 WAT046825 WAT046815

* The same high performance as our conventional Styragel HMW Columns with the added advantage of reducing your solvent consumption by two-thirds. System dead volume must be minimized for maximum column performance.

Calibration Curves for Waters Styragel HMW Series of High-Molecular Weight Columns

Styragel HMW

Styragel HMW 2

Styragel HMW 6E

Styragel HMW 7

Mol

ecul

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t

12 min10 117 8 94 5 6102

103

104

105

106

107

108

109Sample: PolystyreneMobile phase: THFFlow rate: 1 mL/min

307

ULTRASTYRAGEL COLUMNSUltrastyragel™ Preparative Columns provide high-efficiency GPC separations for compound isolation and sample cleanup. Closely related to Styragel GPC Columns, the family of Ultrastyragel Columns provides a two- to three-fold increase in efficiency (plates/meter) that improves separation speed and reduces solvent consumption for preparative isolation. Separations that once required several smaller Styragel Columns can be performed on a single, more efficient, Ultrastyragel Preparative Column.


Ultrastyragel Columns (19 × 300 mm) Ultrastyragel Columns (7.8 × 300 mm)

(mL/min) P/N P/N P/N P/N

Pore Size Effective MW Range Flow Rate Toluene THF Pore Size Effective MW Range Toluene THF

100Å 50–1500 4–10 WAT025866 WAT025859 100Å 50–1500 WAT085500 WAT010570

500Å 100–10,000 4–10 WAT025867 WAT025860 500Å 100–10,000 WAT085501 WAT010571

10^3 Å 200–30,000 4–10 WAT025868 WAT025861

10^4 Å 5000–600,000 4–10 WAT025869 WAT025862

10^5 Å 50,000–4 M 4–10 WAT025870 WAT025863

10^6 Å 200,000–10 M 4–10 WAT025871 WAT025864

Linear 2000–4 M 4–10 WAT025872 WAT025865

HSPgel COLUMNSDesigned for high-speed GPC analysis, the Waters HSPgel™ Column provides an accurate and precise determination of molecular weight, increased sample throughput, and greatly reduced solvent consumption and disposal.

Waters offers these 6.0 × 150 mm columns:

■■ HSPgel HR series, for high-resolution, room-temperature GPC

■■ HSPgel RT series, for routine room temperature GPC

■■ HSPgel HT series for high temperature GPC


High TemperatureWaters HSPgel HT Series5 µm particle sizeAmbient to 180 °C

Room TemperatureWaters HSPgel RT Series3 µm particle sizeAmbient to 80 °C

High ResolutionWaters HSPgel HR Series3 µm particle sizeAmbient to 80 °C

101 102 103 104 105 106 107 108Effective Molecular Weight Range

Effective Molecular Weight Range 101 102 103 104 105 106 107 108

HSPgel HT MB-H

HSPgel HT 1.0

HSPgel HT 2.0

HSPgel HT 2.5

HSPgel HT 3.0

HSPgel HT 4.0

HSPgel HT 5.0

HSPgel HT 6.0

HSPgel HT 7.0

HSPgel HT MB-L

HSPgel HT MB-L/M

HSPgel HT MB-M

HSPgel RT 2.0

HSPgel RT 2.5

HSPgel RT 3.0

HSPgel RT 4.0

HSPgel RT 5.0

HSPgel RT 6.0

HSPgel RT MB-L

HSPgel RT MB-L/M

HSPgel RT MB-M

HSPgel RT MB-H

HSPgel HR 1.0

HSPgel HR 2.0

HSPgel HR 2.5

HSPgel HR 3.0

HSPgel HR 4.0

HSPgel HR MB-L

HSPgel HR MB-M

HSPgel RT 1.0

HSPgel Columns Selection Guide*

HR—High ResolutionRT—Room Temperature

HT—High TemperatureMB—Mixed Bed

L—Low MW RangeM—Medium MW Range

L/M—Low/Medium MW RangeH—High MW Range

*MW ranges for HR and RT are based on polystyrene chain lengths.

309

HSPgel HR Series

The HSPgel HR series is designed for high-resolution, room-temperature, organic polymer GPC. These columns are packed in THF and can be converted once to toluene, dichloromethane, or chloroform.


HSPgel HR Columns, in THF, 3 µm, 6.0 × 150 mm

Description MW Range P/N

HSPgel HR 1.0 100–1000 186001741

HSPgel HR 2.0 500–10,000 186001742

HSPgel HR 2.5 1000–20,000 186001743

HSPgel HR 3.0 2000–60,000 186001744

HSPgel HR 4.0 10,000–400,000 186001745

HSPgel HR MB-L 500–700,000 186001746

HSPgel HR MB-M 1000–4,000,000 186001747

HR—High Resolution, MB—Mixed Bed, L—Low MW Range, M—Medium MW Range.

HSPgel RT Series

The HSPgel RT series is designed for the routine, room-temperature work of organic-polymer GPC. The columns, which are shipped packed in THF, can be converted multiple times, from THF to toluene, chloroform, dichloromethane, DMF, DMSO, etc.


HSPgel RT Columns, in THF, 3 µm, 6.0 × 150 mm


HSPgel RT 1.0 100–1000 186001749

HSPgel RT 2.0 500–10,000 186001750

HSPgel RT 2.5 1000–20,000 186001751

HSPgel RT 3.0 2000–60,000 186001752

HSPgel RT 4.0 10,000–400,000 186001753

HSPgel RT 5.0 25,000–4,000,000 186001754

HSPgel RT 6.0 50,000–10,000,000 186001755

HSPgel RT MB-L 100–10,000 186001757

HSPgel RT MB-L/M 500–400,000 186001758

HSPgel RT MB-M 1000–4,000,000 186001759

HSPgel RT MB-H 5000–10,000,000 186001760

RT—Room Temperature, MB—Mixed Bed, L—Low MW Range, M—Medium MW Range, L/M—Low/Medium MW Range, H—High MW Range.


HSPgel HT Series

The HSPgel HT series is designed for organic GPC conducted at between room temperature and high temperature (180 °C). The columns are shipped packed in either THF or ODCB. The ODCB-packed column should be used for direct conversion to TCB. These columns can withstand multiple solvent switches.


HSPgel HT Columns, in THF, 5 µm, 6.0 × 150 mm HSPgel HT Columns, in ODCB, 5 µm, 6.0 × 150 mm

Description MW Range P/N Description MW Range P/N

HSPgel HT 1.0 100–1000 186001761 HSPgel HT 1.0 100–1000 186001773

HSPgel HT 2.0 500–10,000 186001762 HSPgel HT 2.0 500–10,000 186001774



HSPgel HT 4.0 10,000–400,000 186001765 HSPgel HT 4.0 10,000–400,000 186001777

HSPgel HT 5.0 25,000–4,000,000 186001766 HSPgel HT 5.0 25,000–4,000,000 186001778



HSPgel HT MB-L 100–1000 186001769 HSPgel HT MB-L 100–1000 186001781

HSPgel HT MB-L/M 500–400,000 186001770 HSPgel HT MB-L/M 500–400,000 186001782

HSPgel HT MB-M 1000–4,000,000 186001771 HSPgel HT MB-M 1000–4,000,000 186001783

HSPgel HT MB-H 5000–10,000,000 186001772 HSPgel HT MB-H 5000–10,000,000 186001784

HT - High Temperature, MB - Mixed Bed, L - Low MW Range, M -Medium MW Range, L/M - Low/Medium MW Range, H - High MW Range.

SHODEX COLUMNSWaters is proud to distribute Shodex GPC Columns and accessories. For 30 years, Shodex GPC Columns have been used successfully by scientists worldwide. The following selection of highly-reproducible GPC Columns contains styrene divinylbenzene resins.

K-800 Series (8 × 300 mm)

Ultra-high-efficiency columns designed for high-resolution performance, available in THF, DMF, or chloroform.


Shodex GPC K-800 Columns, in THF, 5 µm, 8 × 300 mm

Description Polystyrene Exclusion Limit P/N

Shodex KF-801 1500 WAT030697

Shodex KF-802 5000 WAT030698

Shodex KF-802.5 20,000 WAT030699

Shodex KF-803 70,000 WAT034100

Shodex KF-804 400,000 WAT034101

Shodex KF-805 4,000,000 WAT034102

Shodex KF-807 200,000,000 WAT034104

Shodex KF-806M (linear) 40,000,000 WAT034105

Shodex KF-G Guard (5 µm, 4.6 × 10 mm) WAT034106

311

Shodex GPC K-800 Columns, in Chloroform, 5 µm, 8 × 300 mm


Shodex K-802.5 20,000 WAT034109

Shodex K-803 70,000 WAT034110

Shodex K-804 400,000 WAT034111

Shodex K-805 4,000,000 WAT034112

Shodex K-G Guard (5 µm, 4.6 × 10 mm) WAT035524

Shodex GPC K-800 Columns, in DMF, 5 µm, 8 × 300 mm


Shodex KD-801 2500 WAT034116

Shodex KD-802 5000 WAT034117

Shodex KD-802.5 20,000 WAT034118

Shodex KD-803 70,000 WAT034119

Shodex KD-804 400,000 WAT034120

Shodex KD-806 40,000,000 WAT034122

Shodex KD-807 200,000,000 WAT034123

Shodex KD-806 M (linear) 40,000,000 WAT034124

Shodex KD-G Guard (5 µm, 4.6 × 10 mm) WAT034125

HFIP-800 Series

These columns have the same high efficiency as the K-series columns shipped in HFIP.

ENVIROGEL HIGH-RESOLUTION GPC CLEANUP COLUMNSThe Envirogel™ High-Efficiency GPC Cleanup Columns remove low volatility, high-molecular-weight interferences, such as lipids and natural resins, from environmental samples, as specified in EPA Method 3640A. In the past, the cleanup procedure for environmental samples was performed on low-efficiency GPC Columns based on packing particle diameters of 37–75 µm (200–400 mesh) Bio-Beads S-X resins. The high efficiency Envirogel GPC Cleanup Columns increase the speed of this process, and simultaneously reduce solvent consumption. For optimum capacity and resolution, a 150 mm column is used in series with the 300 mm column. The use of both the 150 mm and 300 mm column provides maximum loading capacity, while the 300 mm column provides maximum throughput when used alone, plus reduced solvent consumption.


Shodex GPC HFIP-800 Columns, 5 µm, 8 × 300 mm


Shodex HFIP-803 70,000 WAT035605

Shodex HFIP-806M (linear) 40,000,000 WAT035611

Shodex HFP-LG Guard (8 × 50 mm) — WAT035612

Column Optimization

1 2 3 4 5

Collect

Conditions: Column: Envirogel GPC Cleanup, 15 µm, 19 × 150 mm and 19 × 300 mmPart numbers: WAT036555, WAT036554Sample: 2000 µLSolvent: Methylene chlorideFlow rate: 5 mL/minDetection: UV @ 254 nm, 1.5 AUFS

Compounds:1. Corn oil, 62.5 mg/mL2. Bis(2-Ethylhexyl) Phthalate, 2.5 mg/mL3. Methoxychlor, 0.5 mg/mL4. Perylene, 0.05 mg/mL5. Sulfur, 0.2 mg/mL



Envirogel High-Resolution GPC Cleanup Columns

Description Solvent Dimension P/N

Envirogel GPC Cleanup Methylene chloride 19 × 150 mm WAT036555

Envirogel GPC Cleanup Cyclohexane/ethyl acetate 19 × 150 mm 186001915

Envirogel GPC Cleanup Methylene chloride 19 × 300 mm WAT036554

Envirogel GPC Cleanup Cyclohexane/ethyl acetate 19 × 300 mm 186001916

Envirogel GPC Guard Methylene chloride 4.6 × 30 mm 186001913

Envirogel GPC Guard Cyclohexane/ethyl acetate 4.6 × 30 mm 186001914

SEC ColumnsSize-exclusion chromatography (SEC) and gel-filtration chromatography (GFC) are synonymous terms for techniques used to separate macromolecules in aqueous environments according to their hydrodynamic volume. Waters SEC Columns efficiently separate cationic, anionic, and non-ionic macromolecules in many physical, chemical, and biological applications.

ACQUITY APC AQ COLUMNS Designed for aqueous samples, ACQUITY APC AQ Columns are based on hybrid-polymer sub-3-µm particle technology. The advantages of this technology, detailed in the ACQUITY APC XT section on page 301, apply as well to the AQ columns.

10 101 102 103 104 105 106 107


ACQUITY APC AQ 45

ACQUITY APC AQ 125

ACQUITY APC AQ 200

ACQUITY APC AQ 450

ACQUITY APC AQ 900

ACQUITY APC AQ Column Selection Guide

100

1,000

10,000

100,000

1,000,000

10,000,000

0.50 1.00 1.50 2.00

Pol

ysty

rene

Mol

ecul

ar W

eigh

t (D

a)

Elution volume (mL)

ACQUITY APC AQ 450

ACQUITY APC AQ 200

ACQUITY APC AQ 900

ACQUITY APC AQ 125

ACQUITY APC AQ 45

Polystyrene calibration curves for ACQUITY APC AQ Columns.

Column:

313


ACQUITY APC AQ Columns

P/N P/N P/N


30 mm 75 mm 150 mm

45Å 200–5000 1.7 µm 186006972 186006973 186006975

125Å 1000–30,000 2.5 µm 186006977 186006978 186006980

200Å 3000–70,000 2.5 µm 186006982 186006983 186006985

450Å 20,000–400,000 2.5 µm 186006987 186006988 186006990

900Å 300,000–2,000,000 2.5 µm 186007249 186007250 186007251

*All columns are 4.6 mm I.D., maximum temperature limit is 45 °C, columns are shipped dry.



Maximum ResolutionColumns for the analysis of anionic, cationic, and neutral polymers

DPUltrahydrogel D

20Ultrahydrogel 1

Ultrahydrogel 250

Ultrahydrogel 500

Ultrahydrogel 1000

Ultrahydrogel 2000

Ultrahydrogel™ Linear

Higher SpeedColumns for the analysis of anionic, cationic, and neutral polymers

HSPgel AQ MB-H

HSPgel AQ 6.0

HSPgel AQ 5.0

HSPgel AQ 4.0

HSPgel AQ 3.0

el AQ 2.5 AHSPge

Aqueous SEC Column Selection Guide

This chart compares the molecular weight ranges for the specified columns. By connecting two or more columns in series, the effective molecular weight range can be extended to provide coverage for more complex sample analysis.


ULTRAHYDROGEL COLUMNSPacked with hydroxylated, polymethacrylate-based gel, Waters Ultrahydrogel SEC Columns are ideal for analyzing aqueous-soluble samples such as oligomers, oligosaccharides, and polysaccharides. They are likewise well-suited to analyzing cationic, anionic, and amphoteric polymers.

These 7.8 × 300 mm, high-resolution columns offer many advantages over conventional aqueous SEC Columns:

■■ Wide-pH range (2–12)

■■ Compatibility with high concentrations of organic solvents, as much as 20% organic and 50% organic for mobile phases introduced by gradient

■■ Greater flexibility for the mobile phase

■■ Minimal non-size-exclusion effects

Ultrahydrogel Columns Calibration Curves

106

105

104

103

102

4 6 8 10 12 14 min

Mol

ecul

ar W

eigh

t

A B CF

D E

A Ultrahydrogel 120

B Ultrahydrogel 250

C Ultrahydrogel 500D Ultrahydrogel 1000

E Ultrahydrogel 2000

F Ultrahydrogel Linear

Conditions: Column: Ultrahydrogel 200, 500, and 1000 7.8 × 300 mmPart numbers: WAT011525 (250, 6 µm), WAT011530 (500, 10 µm), WAT011535 (1000, 12 µm)Sample: PEG and PEO standardsMobile phase: Distilled waterFlow rate: 1 mL/min

Gelatin Sample

30 min252015

mV

MW = 7500K

MW = 90K

MW = 250K

MW = 480K

Conditions: Column: Ultrahydrogel 250, 500, and 1000, 7.8 × 300 mmPart numbers: WAT011525 (250, 6 µm), WAT011530 (500, 10 µm), WAT011535 (1000, 12 µm)Temperature: 80 °CEluent: Water, pH 7, phosphate bufferFlow rate: 1 mL/minDetection: Waters 410 Differential Refractometer


Ultrahydrogel Columns (7.8 × 300 mm)*

Description Pore Size Particle Size Exclusion Limit P/N

Ultrahydrogel 120 120Å 6 µm 5000 WAT011520

Ultrahydrogel 250 250Å 6 µm 80,000 WAT011525

Ultrahydrogel 500 500Å 10 µm 400,000 WAT011530

Ultrahydrogel 1000 1000Å 12 µm 1,000,000 WAT011535

Ultrahydrogel 2000 2000Å 12 µm 7,000,000 WAT011540

Ultrahydrogel Linear Blend 10 µm 7,000,000 WAT011545

Ultrahydrogel DP* 120Å 6 µm 5000 WAT011550

Ultrahydrogel DNA >2000Å 10 µm 10,000,000 WAT011560

Ultrahydrogel Guard Column N/A 6 µm N/A WAT011565

Ultrahydrogel Guard Column DP* N/A 6 µm N/A WAT011570

*DP = Degree of Polymerization, choice of column when working with glucose oligomers.

315

HSPgel COLUMNSWaters HSPgel™ SEC Columns are optimized for high-speed polymer analysis in aqueous solution. HSPgel Columns reduce solvent consumption, increase throughput, and provide accurate molecular-weight data for any room-temperature analysis. The column dimensions are 6.0 × 150 mm.


HSPgel Columns for High-Speed SEC Analysis*

Description MW Range Solvent Particle Size P/N

HSPgel AQ 2.5 500–2000 Water 4 µm 186001785

HSPgel AQ 3.0 1000–60,000 Water 4 µm 186001786

HSPgel AQ 4.0 10,000–400,000 Water 6 µm 186001787

HSPgel AQ 5.0 50,000–4,000,000 Water 7 µm 186001788

HSPgel AQ 6.0 100,000–10,000,000 Water 9 µm 186001789

HSPgel AQ MB-H 500–10,000,000 Water 9 µm 186001790

*Exclusion limits for AQ series extrapolated from highest MW PEO standard (~900,000).

ACQUITY UPLC PROTEIN SEC COLUMNSACQUITY UPLC Protein SEC Columns are packed with ethylene-bridged hybrid (BEH), diol-coated particle technology. Manufacturers of biotherapeutics and biosimilars can choose the most effective pore size for their application: 125, 200, and 450Å.

Calibration Curves on ACQUITY UPLC Protein BEH SEC, 125Å, 200Å, and 450Å Columns

Protein standards:Temp.: 30 °C Mobile phase: 100 mM sodium phosphate, pH=6.8

MW

100

1000

1,000,000

100,000

10,000

Normalized Retention Volume (Vr/VC)

0.4 1.00.80.6

Uracil (112 Da)Allantoin (158 Da)

Angiotensin frag 1-7 (899 Da)

Aprotinin (6.5 kDa)RNase A (13.7 kDa)

Myoglobin (17 kDa)

Ovalbumin (44 kDa)Conalbumin (75 kDa)

Amyloglucosidase (97 kDa)IgG (150 kDa)

Thyroglobulin (669 kDa)

Blue Dextran

IgM (900 kDa)

ACQUITY UPLC Protein BEH SEC, 450Å, 2.5 µm: 100 K–1500 K Daltons




ACQUITY UPLC Protein BEH SEC 4.6 mm Column

P/N P/N P/N P/N P/N

Pore Size MW Range Particle Size

Column Length

30 mm Guard 150 mm 300 mm 150 mm

w/Standard300 mm

w/Standard

125Å 1K - 80K Da 1.7 µm 186006504 186006505 186006506 176003906 176003907

200Å 10K - 450K Da 1.7 µm 186005793 186005225 186005226 176003904 176003905

450Å 100K - 1500k Da 2.5 µm 186006850 186006851 186006852 176002996 176002997

125Å 1K - 80K Da 1.7 µm — 186008471* — — —

ELSD outlet tubing (0.004" I.D. × 6" length) 430001562

0.005 × 1.75" SEC UPLC connection tubing, 2/pk 186006613

*ACQUITY UPLC Protein BEH SEC 2.1 x 150 mm Column.

For more information on ACQUITY UPLC Protein SEC Columns, refer to page 276.


XBRIDGE PROTEIN BEH SEC COLUMNSXBridge Protein BEH SEC Columns are designed for use on HPLC and UHPLC instrumentation. The 3.5 µm columns are available in 125, 200, and 450Å pore sizes using the same ethylene-bridged hybrid (BEH) particle technology and diol-bonded coating used in Waters UPLC-based SEC columns. This allows you to transfer methods based on laboratory instrumentation and component resolution or sample throughput needs.

Calibration Curves on XBridge Protein BEH SEC, 125Å, 200Å, and 450Å Columns

10

100

1,000

10,000

100,000

1,000,000

10,000,000

0.3 0.4 0.5 0.6 0.7 0.8 0.9 1

MW

Normalized Retention Volume (V0/VC)




Uracil (112 Da)

Aprotinin (6.5 KDa) RNase A (14 KDa) Myoglobin (17 KDa)

Ovalbumin (44 KDa) Conalbumin (75 KDa)

Amyloglucosidase (97 KDa) IgG (150 KDa)

Thyroglobulin (669 KDa)

IgM (900 KDa)


XBridge Protein BEH SEC, 7.8 mm I.D. Columns*

P/N P/N P/N


30 mm 75 mm 150 mm

125Å 1K - 80K 3.5 µm 176003591 176003592 176003593

200Å 10K - 450K 3.5 µm 176003594 176003595 176003596

450Å 100K - 1500k 3.5 µm 176003597 176003598 176003599

Straight Connection Tubing and End-fittings for XBridge Protein BEH SEC Column WAT022681

U-Bend Connection Tubing and End-fittings for XBridge Protein BEH SEC Column WAT084080

* All columns and guards include standards mix. SEC Protein Standards are matched to the pore size of the column.

317

PROTEIN-PAK SIZE-EXCLUSION HPLC COLUMNSProtein-Pak Packings are based on a 10 µm diol-bonded silica and are available in a selection of pore sizes and column configurations.

The Protein-Pak Size-Exclusion Columns can be expected to resolve proteins that differ in molecular weight by a factor of two and to distinguish proteins differing by as little as 15% in molecular weight. The degree of resolution is more dependent on the sample mass and volume than the interaction between the sample and the stationary phase. Ideally, there should be no interaction between the stationary phase and the sample molecules. Secondary interactions are most often ionic and can, therefore, be reduced by increasing the ionic strength of thee mobile phase. Typical, salt concentrations range to 0.2-0.5 M NaCl. It may also be useful in some cases to consider adding 10-20% methanol to eliminate hydrophobic and other hydrogen-bonding interactions.

For more information on XBridge Protein BEH SEC Columns, refer to page 280.


Protein-Pak SEC HPLC Columns and Guards

Steel Column Dimension MW Range P/N

Protein-Pak 60 7.8 x 300 mm 1,000–20,000 WAT085250

Protein-Pak 60 19 x 300 µm 1,000–20,000 WAT025830

Protein-Pak 125 7.8 x 300 mm 2,000–80,000 WAT084601

Protein-Pak 125 19 x 300 mm 2,000–80,000 WAT025831

Protein-Pak 300SW 7.5 x 300 mm 10,000–300,000 WAT080013

Protein-Pak 125 Sentry Guard Column, 3.9 x 20 mm, 2/pk (requires holder) 186000926

Sentry Universal Guard Column Holder WAT046910

PROTEIN STANDARDSEach standard contains proteins selected for ACQUITY UPLC and XBridge Protein BEH SEC Columns. Use these standards for purposes of quality control, to test an HPLC or UPLC column, and to monitor column performance over time.


BEH SEC Column Protein Standards

Description P/N

BEH125 SEC Protein Standard Mix 186006519

A mix of 4 proteins; Thyroglobulin, ovalbumin, ribonuclease A and uracil


A mix of 5 proteins; Thyroglobulin, IgG, BSA, Myoglobin, Uracil


A mix of 5 proteins; Thyroglobulin, IgG, BSA, Myoglobin, Uracil


Solvent GuideThe following graphic is a guide for eluents.

Polymer Class Eluent

Polyethylene oxidePolyethylene glycolPolysaccharides, pullulans Dextrans Celluloses (water-soluble)Polyvinyl alcohol Polyacrylamide

0.10 M Sodium nitrateNeutral

Polyacrylic acid Polyalginic acid/alginates Hyaluronic acid Carrageenan

0.10 M Sodium nitrateAnionic

DEAE dextran Polyvinylamine

Polyepiamine

n-Acetylglucosamine

0.80 M Sodium nitrate

0.10% TEA

0.10 M TEA/1% Acetic acid

Cationic

Cationic

Cationic

Polyethyleneimine Poly(n-methyl-2-vinyl pyridinium) I salt

Lysozyme Chitosan

Polylysine

Peptides

0.50 M Sodium acetate/0.50 M Acetic acid

0.50 M Acetic acid/0.30 M Sodium sulfate

5% Ammonium biphosphate/3% Acetonitrile (pH = 4.0)

0.10% TFA/40% Acetonitrile

Cationic, hydrophobic




Polystyrene sulfonate Lignin sulfonate

Anionic, hydrophobic

Collagen/gelatin Amphoteric

Polyvinyl pyrrolidone

80:20 0.10 M Sodium nitrate/Acetonitrile

Neutral, hydrophobic

Aqueous SEC Solvent Selection Guide

319

Polyisobutylene

PolybutyleneChlorinated rubberPolybutadienePolyisoprenePolydimethylsiloxane

Chlorinated polyethylenePolyethylene–EthylacrylatePolyethylene–VinylacetonePolyethylene–Methacrylic acidPolyphenyleneoxidePoly-4-methylpentene(1)Polyethylene

Ultra-high Molecular Weight PolyethylenePolypropylene

PolyetheretherketonePolyetherketone

Polycarbonate

Polyglycolic acid

Acrylonitrile–MethylmethacrylateCellulose acetateCellulose acetate–ButyrateCellulose acetate–PropionateCellulose nitrate Cellulose propionateCellulose triacetateDiallyl phthalateEthyl celluloseEpoxyPolyester alkydPolybutene(1)Polybutadiene–StyrenePhenol–FormaldehydePhenol–FurfuralPolymethylmethacrylatePolypropyleneglycolPolystyrenePolysulfonePolyvinylacetatePolyvinylbutyralPolyvinylchloridePolyvinylchloride–AcetatePolyvinyldienechloridePolyvinylformalPolystyrene acrylonitrilePolystyrene–AlphamethylstyrenePolyester thermosetPhenolicsRosin acidsPolyglycolic acid

Melamine–FormaldehydeNylon (all types)Polybutylene–TeraphthalatePolyethylene–Teraphthalate

Poly acrylonitrileABS (Acrylonitrile–Butadiene–Styrene)ASA (Acrylic–Styrene–AcrylonitrileABA (Acrylonitrile–Butadiene–Acrylate)Carboxymethyl celluloseABS/PolycarbonatePolybutadiene–AcrylonitrilePolyurethane

PolyacetalPolyoxymethylene

PolyimidePolyamide–imidePolyetherimidePolyethersulfonePolyvinyldienefluoride

Polyfuran–Formaldehyde

Polymer

Toluene

Toluene/75 °C

TCB/135–160 °C

TCB/135–160 °C

Phenol/TCB 1:1/145 °C

Methyl chloride

gamma-Butyl lactone

THF/40 °C

Hexafluoroisopropanol + 0.075 MSodium trifluoroacetate/55 °Corm-Cresol + 0.05 m LiBr/100 °C

DMF + 0.05 m LiBr/85 °C

DMF + 0.05 m LiBr/145 °C

N-Methyl pyrrolidone+ 0.05 m LiBr/100 °C

Dimethylacetamide/60 °C

GPC Solvent

Waters Styragel Columns

shipped in Toluene


shipped in THF


shipped in DMF

Shipping Solvent

Non-Aqueous GPC Solvent Selection Guide


Calibration StandardsWaters offers a selection of well-characterized polymer standards for calibration. The offering includes kits as well as individual standards. The standards are available for aqueous and non-aqueous applications.

ACQUITY APC CALIBRATION STANDARDSACQUITY APC Calibration Standards match the molecular-weight range of the ACQUITY APC XT Columns. These kits eliminate the need to manually prepare custom calibration mixes because they provide the correct number of data points for the targeted molecular-weight range. In addition, they reduce, by 3–5 times, the ACQUITY APC System’s calibration time. With reduced calibration time, calibrations can be carried out on a more frequent basis, increasing confidence in the accuracy of results.

AU

0.000

0.035

0.070

0.105

0.140

2.80 3.50 4.20 4.90 5.60 6.30 min

V1 = 130,000 +21,500 + 6,540 + 1,250V2 = 66,000 + 15,700 + 3,470 + 474V3 = 35,500 + 9,130 + 2,280 + 266

Medium MW standards:MW: 266–130,000Columns: 45Å + 125Å + 200Å 150 mm each THF at 0.8 mL/min 10 µL PDA @ 254 nm

The ACQUITY APC Calibration Standards are available in both polystyrene and polymethyl methacrylate, configured as low, middle, and high-molecular-weight calibration kits. Also available are method development kits, which include the full separation range of the three kits combined.


ACQUITY APC Calibration Standards


ACQUITY APC Polystyrene Low MW Calibration Kit

3 vials containing different MW ranges, Qty. 10 of each vial266–15,000 186007539

ACQUITY APC Polystyrene Middle MW Calibration Kit


ACQUITY APC Polystyrene High MW Calibration Kit

3 vials containing different MW ranges, Qty. 10 of each vial266–2,500,000 186007541

ACQUITY APC Polystyrene Method Development MW Calibration Kit

9 vials containing 1 vial each of the low, middle, and high polystyrene kits266–2,500,000 186007542

ACQUITY APC Polymethyl Methacrylate Low MW Calibration Kit


ACQUITY APC Polymethyl Methacrylate Middle MW Calibration Kit


ACQUITY APC Polymethyl Methacrylate High MW Calibration Kit

3 vials containing different MW ranges, Qty. 10 of each vial202–1,600,000 186007545

ACQUITY APC Polymethyl Methacrylate Method Development MW Calibration Kit202–1,600,000 186007546

9 vials containing 1 vial each of the low, middle, and high polymethyl methacrylate kits

321

Non-Aqueous GPC Standards Guide

Reference materials for the analysis of organic soluble polymers



101 102 103 104 105 106 107

101 102 103 104 105 106 107

Polystyrene ReadyCal

Polybutadiene

Polyisoprene

Polymethylmethacrylate

Polymethylmethacrylate

READYCAL STANDARDSA ReadyCal Kit allows quick and accurate preparation of a multi-point calibration curve without the need to weigh chemicals. Each vial contains a polymer mix that spans a molecular-weight range, to provide baseline resolution of each component. Simply add solvent directly to the vial and mix.


ReadyCal Standards

Description* P/N

Polystyrene ReadyCal Standards 4 mL Kit

A complete kit of ready-to-use polystyrene calibration standards. Kit contains thirty, 4 mL autosampler vials which contain four polystyrene standards per vial. There are three separate molecular weight ranges in each kit, ten units of each range. Range is from 400 to 2,000,000 Da.

WAT058930

Polystyrene ReadyCal Standards 2 mL Kit

A complete kit of ready-to-use polystyrene calibration standards. Kit contains thirty, 2 mL autosampler vials which contain four polystyrene standards per vial. There are three separate molecular weight ranges in each kit, ten units of each range. Range is from 400 to 2,000,000 Da.

WAT058931

*Values listed are approximate molecular weights.


POLYMER SPECIFIC CALIBRATION STANDARDSTailored specifically for different types of polymer analysis, these calibration standards provide a quick and reliable reference to known molecular-weight ranges. Polymer type and MW ranges appear in the table.


Polymer Specific Calibration Standards

Description* P/N

Polybutadiene Standards Kit

0.5 g/vial polybutadiene at each molecular weight: 1000, 3000, 7000, 10,000, 30,000, 70,000, 100,000, 300,000, 700,000, 1,000,000WAT035709

Polyisoprene Standards Kit

0.5 g/vial polyisoprene at each molecular weight: 1000, 3000, 10,000, 30,000, 70,000, 100,000, 300,000, 500,000, 1,000,000, 3,000,000WAT035708

Polymethylmethacrylate Low MW Standards Kit

0.5 g/vial polymethylmethacrylate at each molecular weight: 1000, 1700, 2500, 3500, 5000, 7000, 10,000, 13,000, 20,000, 30,000WAT035707

Polymethylmethacrylate Mid MW Standards Kit

0.5 g/vial polymethylmethacrylate at each molecular weight: 2400, 9500, 31,000, 52,000, 100,000, 170,000, 270,000, 490,000, 730,000, 1,000,000

WAT035706

Polystyrene Low-Mid MW Standards Kit

10 g/vial polystyrene at each molecular weight: 400, 530, 950 5 g/vial polystyrene at each molecular weight: 2800, 6400, 10,000, 17,000, 43,000, 110,000, 180,000

WAT011588

Polystyrene Mid-High MW Standards Kit

5 g/vial polystyrene at each molecular weight: 430,000, 780,000 1 g/vial polystyrene at each molecular weight: 1,300,000, 2,800,000, 3,600,000, 4,300,000, 5,200,000, 6,200,000, 8,400,000, 20,000,000

WAT011610

Polystyrene Low MW Standards Kit

0.5 g/vial polystyrene at each molecular weight: 580, 950, 1200, 1800, 2470, 3770, 5100, 7600, 12,500, 17,000WAT034208

Polystyrene Mid MW Standards Kit

0.5 g/vial polystyrene at each molecular weight: 1200, 3250, 10,200, 28,000, 68,000, 195,000, 490,000, 1,080,000, 1,750,000, 2,750,000WAT034209

Polystyrene High MW Standards Kit

0.5 g/vial polystyrene at each molecular weight: 45,000, 1,270,000, 2,300,000, 3,260,000, 4,340,000, 8,000,000, 15,000,000WAT034210


323

INDIVIDUAL MW STANDARDSIn many cases, a single calibration standard can verify the molecular weight of a sample-mixture component, making its identification simple and straightforward.


Individual MW Standards

Description* P/N Description* P/N

Polystyrene Standard 400

10 g/vial polystyrene, 400 MWWAT011590

Polystyrene Standard 430,000

5 g/vial polystyrene, 430,000 MWWAT011612







Polystyrene Standard 1,300,000

1 g/vial polystyrene, 1,300,000 MWWAT011616






























SEC CALIBRATION STANDARDSWaters SEC Calibration Standards are precisely formulated to determine accurate molecular weight and conveniently packaged to minimize errors in SEC calibration methods. The fully traceable aqueous-based polymer kits simplify routine calibration procedures that improve workflow and increase productivity.

Aqueous SEC Standards Guide

Standards for the analysis of anionic, cationic, and neutral polymers



Polyacrylate

Pullulan

Dextran

Polyethylene Oxide

Polyethylene Glycol

This chart may be used to determine the appropriate component standard and corresponding molecular weight range.


Full-Range Calibration Standards

These standards kits provide an accurate calibration range for determining the molecular weight of common water-soluble polymers. The kits contain a series of well-characterized standards of a specified polymer type and include certificates that list component ranges and concentrations.


Full-Range Calibration Standards for SEC

Description* P/N

Polyacrylic Acid Standards Kit

250 mg/vial polyacrylic acid at each molecular weight: 1000, 3000, 7000, 15,000, 30,000, 70,000, 100,000, 300,000, 700,000, and 1,000,000

WAT035714

Polyethylene Glycol Standards Kit

1.0 g/vial polyethylene glycol at each molecular weight: 100, 200, 400, 600, 1000, 1500, 4300, 7000, 13,000, and 22,000

WAT035711

Polyethylene Oxide Kit

500 mg/vial polyethylene oxide at each molecular weight: 24,000, 40,000, 79,000, 160,000, 340,000, 570,000, and 850,000

WAT011572

Dextrans Standard

500 mg/vial dextrans at each molecular weight: 5000, 12,000, 24,000, 48,000, 148,000, 273,000, 410,000, and 750,000

WAT054392

Pullulan Kit

200 mg/vial pullulan at each molecular weight: 5000, 10,000, 20,000, 50,000, 100,000, 200,000, 400,000, and 800,000

WAT034207


Individual Calibration Standards

In many cases, a single calibration standard can verify the molecular weight of a sample-mixture component, making its identification simple and straightforward.


Individual Calibration Standards for SEC

Description* P/N

Polyethylene Oxide Standard 24,000 WAT011574








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